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1.
Figure 3

Figure 3. Levels of IGF1R during eosinophil differentiation culture from the miR-223+/+ and miR-223-/- mice. From: MiR-223 deficiency increases eosinophil progenitor proliferation.

(A) Western blot showing levels of pre-IGF1R and IGF1R in eosinophil differentiation culture derived from miR-223+/+ and miR-223-/- mice from day 4 to day 14. GAPDH is shown as a loading control.

Thomas X. Lu, et al. J Immunol. ;190(4):1576-1582.
2.
Figure 1

Figure 1. MiR-223 is induced during eosinophil differentiation. From: MiR-223 deficiency increases eosinophil progenitor proliferation.

(A) Schematic of ex vivo bone marrow derived eosinophil culture. (B) Purity of cultured eosinophils after 14 days. Eosinophils are identified as CCR3+Siglec-F+ cells. (C) Levels of miR-223 during the eosinophil differentiation culture. N = 3 per group; data are represented as mean ± S.E.M.

Thomas X. Lu, et al. J Immunol. ;190(4):1576-1582.
3.
Figure 2

Figure 2. Proliferation of eosinophil progenitor cells and morphology of mature eosinophils from miR-223+/+ and miR-223-/- cultures during the ex vivo eosinophil differentiation culture. From: MiR-223 deficiency increases eosinophil progenitor proliferation.

(A) Total cell number of eosinophils in cultures derived from miR-223+/+ & miR-223-/- mice, determined by cell counting using a hemacytometer. ** p < 0.01; N = 6 per group; data are represented as mean ± S.E.M. (B) Morphology of miR-223+/+ & miR-223-/- cultured eosinophils at day 14 determined by Diff-Quik staining Magnification: 400x.

Thomas X. Lu, et al. J Immunol. ;190(4):1576-1582.
4.
Figure 5

Figure 5. Analysis of eosinophil progenitor culture expression of CCR3. From: MiR-223 deficiency increases eosinophil progenitor proliferation.

(A) CCR3 expression at day 8, day 10 and day 12 of the eosinophil progenitor culture were determined in miR-223-/- cultures compared to miR-223+/+ cultures by qRT-PCR normalized to HPRT1. N = 3 per group; data are represented as mean ± S.E.M. (B) Levels of CCR3 expression during eosinophil differentiation culture determined by FACS staining of surface CCR3 and Siglec F levels. Mature eosinophils are identified as CCR3+Siglec-F+ cells.

Thomas X. Lu, et al. J Immunol. ;190(4):1576-1582.
5.
Figure 6

Figure 6. Heatmap of differentially regulated genes between miR-223+/+ and miR-223-/- eosinophil progenitor cultures at day 8 and day 12 with their associated top biological functions. From: MiR-223 deficiency increases eosinophil progenitor proliferation.

(A) Heat map of differentially regulated genes at day 8 of the eosinophil differentiation culture. Red: up-regulated in miR-223-/- eosinophil progenitor culture compared to miR-223+/+ eosinophil progenitor culture; blue: down-regulated in miR-223-/- eosinophil progenitor culture compared to miR-223+/+ eosinophil progenitor culture. (B) Functional enrichment analysis of differentially regulated genes in the eosinophil progenitor cultures at day 8. The networks are shown as Cytoscape graph networks generated from ToppCluster network analysis. (C) Heat map of differentially regulated genes at day 12 of the eosinophil differentiation culture. Red: up-regulated in miR-223-/- eosinophil progenitor culture compared to miR-223+/+ eosinophil progenitor culture; blue: down-regulated in miR-223-/- eosinophil progenitor culture compared to miR-223+/+ eosinophil progenitor culture. (D) Ingenuity analysis of the most significant biological functions represented by the differentially regulated genes between miR-223+/+ and miR-223-/- eosinophil progenitor cultures at day 12.

Thomas X. Lu, et al. J Immunol. ;190(4):1576-1582.
6.
Figure 4

Figure 4. The effect of IGFR1 inhibitor on proliferation of eosinophil progenitors. From: MiR-223 deficiency increases eosinophil progenitor proliferation.

(A) Bone marrow derived eosinophils from miR-223+/+ and miR-223-/- mice were treated with 2 μm picropodophyllin (PPP, an IGF1R inhibitor) or an equivalent volume of DMSO. Cell proliferation is measured by cell counting using a hemacytometer. (B) Levels of IGF1R expression in miR-223+/+ and miR-223-/- cells after 2-day treatment with PPP. GAPDH is used as a loading control. (C) A dose response study of different concentrations of PPP on the proliferative response of eosinophil cultures derived from miR-223+/+ and miR-223-/- mice. N = 3 per group; data are represented as mean ± S.E.M.

Thomas X. Lu, et al. J Immunol. ;190(4):1576-1582.

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